Introduction
The HiPure Plasmid DNA Mega Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 10 mg high copy number plasmid DNA can be purified from 500 mL overnight culture.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation up to 10mg endotoxin-free plasmid DNA from 500ml bacterial culture |
| Applications | Cell transfection, animal injection, etc. |
| Purification method | Mega spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Conventional plasmid, plasmid less than 30KB |
| Sample amount | High copy plasmid: 500ml culture medium |
| Yield | 10mg |
| Elution volume | ≥0.8ml |
| Time per run | ≤70 minutes |
| Liquid carrying volume per column | 50ml |
| Binding yield of column | 10mg |
Principle
The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
- High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
- Fast – silica gel column purification is much faster than ion exchange
- High yield – up to 10mg plasmid can be binded in one column
- Low endotoxin – the obtainedplasmid can be directly used for cell transfection and animal injection, etc.
Kit Contents
| Contents | P111602 | P111603 |
| Purification Times | 10 Preps | 50 Preps |
| RNase A | 60 mg | 2 x 150 mg |
| Buffer E1 | 220 ml | 2 x 550 ml |
| Buffer E2 | 220 ml | 2 x 550 ml |
| Buffer E3 | 220 ml | 2 x 550 ml |
| Buffer E4 | 220 ml | 2 x 550 ml |
| Buffer E5 | 120 ml | 550 ml |
| Buffer PW2 | 25 ml | 2 x 100 ml |
| Elution Buffer | 30 ml | 120 ml |
| HiPure EF Maxi Columns | 10 | 50 |
| Lysate Clear Maxi Syringe | 10 | 50 |
| 50 ml Collection Tubes | 20 | 100 |
Storage and Stability
The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve.
Purchase Guide
| Name | CAT NO | Sample amount | Column type | Elution volume | Yield | Time per run | Centrifuge requirements |
| HiPure Plasmid EF Mini Kit | P1154 | 10-15ml | 1.5ml column | ≥75μl | 10–70µg/15ml | ≤40 minutes | small centrifuge |
| HiPure Plasmid EF 96 Kit | P1157 | 1-5ml | 96 well plate | ≥75μl | 30µg/1ml | ≤60 minutes | 96 well plate centrifuge |
| HiPure Plasmid EF Midi Kit | P1231 | 30-50ml | 15ml column | ≥0.4ml | 10-500μg/50ml | ≤60 minutes | 15ml centrifuge |
| HiPure Plasmid EF Maxi Kit | P1156 | 100-200ml | 50ml column | ≥0.8ml | 100–1500µg/200ml | ≤60 minutes | 50ml centrifuge |
| HiPure Plasmid EF Mega Kit | P1116 | 500ml | 50ml column | ≥0.8ml | 10mg/500ml | ≤70 minutes | 50ml centrifuge |
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